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1.
为了实现二斑叶螨Tetranychus urticae Koch对阿维菌素抗药性的快速检测,本研究建立了一种叶螨对阿维菌素抗药性的快速分子检测技术,即酶切扩增多态性序列(cleaved amplified polymorphic sequences,CAPS)标记技术,检测二斑叶螨谷氨酸门控氯离子通道(Glu Cl3)基因片段上G326E的突变频率,并采用琼脂浸叶法测定田间5个种群对阿维菌素的抗药性。结果表明,北京密云、顺义、昌平、浙江宁波和海南吉阳5个种群对阿维菌素的抗性倍数分别达1 155.63、1 317.10、844.19、314.95和1 799.69倍,均为极高抗性水平;CAPS检测发现我国二斑叶螨抗性基因存在G326E位点突变,但在供试的5个不同种群中突变频率变异很大,其中北京昌平种群G326E突变频率高达90.00%,密云和顺义种群均为26.67%,浙江宁波种群为5.00%,而海南吉阳种群不存在该位点的突变。表明该CAPS标记还需与其它抗性机制或检测技术结合来实现叶螨对阿维菌素抗性的早期检测和预警。 相似文献
2.
通过解毒酶活性测定与钠离子通道基因片段的克隆、测序,研究了南京小菜蛾对拟除虫菊酯产生高水平抗性的生化和分子机制。结果表明,南京抗性种群的酯酶和多功能氧化酶O-脱甲基活性分别是敏感种群的1.06和1.16倍,无显著差异;以2,4二硝基氯苯(CDNB)和1,2-二氯-4-硝基苯(DCNB)为底物,南京抗性种群的谷胱甘肽S-转移酶活性分别是敏感种群的0.68倍和1.03倍;进一步利用聚合酶链式反应(PCR)分别从敏感和抗性小菜蛾基因组DNA中扩增出了位于钠离子通道结构域IIS6的232bp和248bp的DNA片段;与敏感种群相比,南京抗性种群钠离子通道基因存在一个保守性点突变,即C突变为T,并导致亮氨酸突变为苯丙氨酸,表明神经不敏感性是南京小菜蛾对拟除虫菊酯产生高水平抗性的主要机制。 相似文献
3.
浙江省二化螟不同种群和大螟对三唑磷的敏感性研究 总被引：4，自引：0，他引：4
于2004年用点滴法测定了浙江杭州地区(富阳和余杭)和嘉兴地区(平湖、秀城和秀洲)二化螟、杭州富阳大螟等种群4龄幼虫对三唑磷的敏感性,以及各种群体内与抗药性相关酶系(多功能氧化酶、谷胱甘肽S-转移酶、酯酶和乙酰胆碱酯酶等)的比活力。结果证实上述地区二化螟种群对三唑磷产生了21.1~218.8倍的抗性,其中以富阳和平湖种群的抗性水平较高。二化螟体内多功能氧化酶活性与三唑磷抗性水平呈显著正相关,而乙酰胆碱酯酶活性则呈显著负相关,表明二化螟对三唑磷的抗性可能与多功能氧化酶活性增强和乙酰胆碱酯酶敏感性降低有关。大螟富阳种群对三唑磷的敏感性明显高于二化螟富阳种群,其抗性机制还有待研究。 相似文献
4.
灰飞虱对杀虫剂的抗性分子机制研究进展 总被引：1，自引：0，他引：1
灰飞虱是中国长江流域和黄淮地区重要的农业害虫,由于杀虫剂的广泛与大量使用,已导致其对多种杀虫剂产生了抗性。深入研究其抗药性分子机制,可为灰飞虱抗性的快速检测和治理提供重要理论基础。文章总结了灰飞虱对毒死蜱、吡虫啉、溴氰菊酯、噻嗪酮、氟虫腈和乙虫腈等杀虫剂的抗性分子机制研究进展,主要包括抗性相关解毒酶和转运蛋白基因的筛选与功能验证,以及靶标位点突变等重要研究成果,指出该研究领域当前存在的问题主要有抗性基因的功能验证及调控路径、抗性新基因的鉴定及交互抗性和多重抗性机制不明确等,并展望了其未来发展方向,认为:可利用CRISPR/Cas9基因编辑技术验证抗性基因功能;可将转录组测序结合生物信息学手段用于鉴定新抗性基因及抗性调控基因,以探明交互抗性和多重抗性机制;应深入至蛋白组学水平探讨抗性机制;需开发配套的高效田间施药技术,以达到杀虫剂减施增效的目的。 相似文献
5.
昆虫对拟除虫菊酯类杀虫剂的代谢抗性机制研究进展 总被引：1，自引：1，他引：0
随着拟除虫菊酯类杀虫剂在卫生和农业害虫防治中的广泛应用,昆虫对此类杀虫剂产生抗性的报道越来越多。目前已明确昆虫对拟除虫菊酯类杀虫剂的抗性机制包括表皮穿透率下降、靶标抗性以及代谢抗性,其中代谢抗性机制较为普遍,而且其与昆虫对多种杀虫剂的交互抗性关系密切。目前,随着基因组、转录组以及蛋白质组学等新技术的发展及应用,昆虫对拟除虫菊酯类杀虫剂的代谢抗性机制研究也取得了很多新进展。昆虫体内细胞色素P450酶（P450s）、羧酸酯酶（CarE）及谷胱甘肽S-转移酶（GSTs）等重要解毒酶系的改变均与昆虫对拟除虫菊酯类杀虫剂的代谢抗性有关,其中这3类解毒酶的活性及相关基因表达量的变化是昆虫对此类杀虫剂产生代谢抗性的主要原因。明确昆虫对拟除虫菊酯类杀虫剂的代谢抗性机制,对合理使用此类杀虫剂及延缓抗药性的产生均具有重要意义。本文在总结拟除虫菊酯类杀虫剂代谢路径及相关生物酶研究概况的基础上,综述了近年来有关昆虫对此类杀虫剂代谢抗性机制研究的主要进展。 相似文献
6.
新型双酰胺类杀虫剂已广泛用于保障水稻生产,而二化螟作为危害水稻生产的钻蛀性害虫,已经对该类杀虫剂产生了抗性,明确该类杀虫剂抗性分子机制,可为二化螟抗性快速检测和绿色防控提供技术支撑。本文在总结二化螟对双酰胺类杀虫剂抗性现状的基础上,重点综述了近年来有关其抗性分子机制研究的进展,主要包括解毒酶和转运蛋白基因过表达介导的代谢抗性,以及鱼尼丁受体基因突变介导的靶标抗性;指出了该研究领域在抗性分子检测、抗性新基因鉴定、抗性基因调控网络和多重抗性机制等方面存在的问题,并展望了其发展方向,认为：利用高通量测序技术检测害虫种群抗药性;利用多组学技术鉴定新抗性基因及调控网络,以探明多重抗性机制;将反向遗传学工具放射性配基结合及电生理技术深入验证抗性基因功能;需开发靶向抗性基因的dsRNA转基因作物、纳米农药及选择性新型化学杀虫剂,以达到杀虫剂减施增效的目的。 相似文献
7.
棉铃虫Bt抗感种群间数种解毒酶和中肠蛋白酶活性的比较 总被引：12，自引：3，他引：12
比较Bt抗性种群和敏感种群棉铃虫解毒酶和中肠蛋白酶的变化，测定两种群的α-乙酸萘酯酶、乙酰胆碱酯酶、谷胱甘肽-S-转移酶的酶活力，比较α-乙酸萘酯酶和乙酰胆碱酯酶的Km和Vmax值，并采用特异性酶抑制剂进行鉴定，结果发现，抗性种群棉铃虫的乙酰胆碱酯酶活力、Km和Vmax值都 显著升高，甲基对氧磷对抗性种群棉铃虫乙酰胆碱酯酶的抑制程度比敏感种群明显霸产加，抗生种群5和敏感种群总蛋白酶活力差异不显著，弱碱性类胰蛋白酶和类凝乳蛋白酶活力差异也不显著，但抗性种群3龄幼虫的强碱性类胰蛋白酶活图形显著高于敏感种群。 相似文献
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棉蚜Aphis gossypii Glover是棉花生产中最严重的害虫之一。长期以来防治棉蚜主要依赖于化学杀虫剂,其中新烟碱类杀虫剂扮演着十分重要的角色,但由于其长期、大量的使用,棉蚜已对该类杀虫剂产生了较高水平的抗性,严重影响了对棉蚜的防治效果。抗性机制研究表明,棉蚜对新烟碱类杀虫剂产生抗性的机制主要涉及解毒代谢能力增强和靶标敏感性下降。细胞色素P450、羧酸酯酶、谷胱甘肽S-转移酶、UDP-葡糖基转移酶等解毒酶基因过量表达介导的解毒代谢增强和烟碱型乙酰胆碱受体β1亚基突变引起的靶标敏感性下降是棉蚜对新烟碱类杀虫剂产生抗性的关键。针对我国棉蚜对新烟碱类杀虫剂抗性问题突出的现状,为更好地利用该类药剂防治棉蚜,亟需根据棉蚜抗性机制制定合理的抗性治理策略。本文将从棉蚜对新烟碱类杀虫剂的抗性现状、抗性机制以及抗性治理策略等方面进行综述,以期为抗性棉蚜的科学治理提供参考。 相似文献
10.
《中国生物防治学报》2017,(6)
为在小菜蛾防治中合理使用化学杀虫剂和生物农药Bt,探究了小菜蛾对化学农药的抗药性与其对Bt杀虫蛋白Cry1Ac的敏感度之间的关系。采用浸叶法测定了2个抗性背景不同的小菜蛾种群对9种常用杀虫剂和Cry1Ac蛋白的敏感度,通过比较敏感度差异,初步探索了小菜蛾对化学农药的抗药性和其对Cry1Ac蛋白敏感度之间的关系。结果表明:采自广东博罗县田间的小菜蛾种群(GDBL)对除Cry1Ac之外的9种常用杀虫剂,均已产生了不同程度的抗药性;而北京室内小菜蛾种群(BJNK),虽然对高效氯氰菊酯、阿维菌素、茚虫威、多杀菌素、溴虫腈具有抗性,对Cry1Ac在内的其他5种杀虫剂仍比较敏感。可见,对多种不同类型的杀虫剂产生了不同程度抗性的两个小菜蛾种群,对Cry1Ac蛋白仍处于敏感状态。由于小菜蛾对化学杀虫剂的抗性机制与其对Cry1Ac的抗性机制不同,小菜蛾对化学杀虫剂的抗药性与其对Cry1Ac的抗药性之间不存在交互抗性。因此,在田间可以通过加强Bt杀虫剂的使用来提高对多种化学杀虫剂产生抗性的小菜蛾的防治效果。 相似文献
11.
为研究CYP6家族P450基因在小菜蛾Plutella xylostella代谢氯虫苯甲酰胺中的作用,利用浸叶法测定不同小菜蛾种群3龄幼虫对氯虫苯甲酰的抗性水平,采用实时荧光定量PCR(quantitative realtime PCR,q RT-PCR)方法分析CYP1v3、CYP1v4、CYP6B6和CYP6f这4种P450基因在小菜蛾不同抗性种群体内的表达差异及杀虫剂的短期诱导效应,并通过RNA干扰技术沉默4种P450基因后分析小菜蛾对氯虫苯甲酰胺的敏感性。结果显示,4种P450基因在中等抗性水平小菜蛾种群体内高表达;氯虫苯甲酰胺处理可诱导4种基因显著上调表达。分别沉默4种P450基因后,处理组小菜蛾的P450酶活力显著下降37.60%～54.82%,且处理组小菜蛾的死亡率显著高于对照组;同时沉默4种基因处理组的小菜蛾P450酶活力显著低于其他各处理组。表明4种P450基因可能同时参于小菜蛾对氯虫苯甲酰胺的代谢。 相似文献
12.
Shoji Sonoda 《Pest management science》2010,66(5):572-575
BACKGROUND: The pyrethroid resistance of the diamondback moth Plutella xylostella (L.) is conferred by increased gene expression of cytochrome P450 to detoxify the insecticide and/or through gene mutation of the sodium channel, which makes the individual insensitive to pyrethroids. However, no information is available about the correlation between the increased metabolic detoxification and the target insensitivity in pyrethroid resistance. RESULTS: Frequencies of pyrethroid‐resistant alleles (L1014F, T929I and M918I) and two resistance‐related mutations (A1101T and P1879S) at the sodium channel and expression levels of the cytochrome P450 gene CYP6BG1 were examined individually using laboratory and field strains of P. xylostella. Real‐time quantitative PCR analysis using the laboratory strains revealed that levels of larval expression of the resistant strain, homozygous for the pyrethroid‐resistant alleles other than the M918I, are significantly higher than those of the susceptible strain. In the field strains, the expression levels in insects having the same resistant alleles as those of the resistant strains varied greatly among individuals. The expression levels were not significantly higher than those in the heterozygotes. CONCLUSION: Significant correlation between the target insensitivity and the increased metabolic detoxification in pyrethroid resistance of P. xylostella was observed in the laboratory but not in the field. Copyright © 2010 Society of Chemical Industry 相似文献
13.
Decreased acetylcholinesterase (AChE) sensitivity and metabolic detoxification mediated by glutathione S-transferases (GSTs) were examined for their involvement in resistance to acephate in the diamondback moth, Plutella xylostella. The resistant strain showed 47.5-fold higher acephate resistance than the susceptible strain had. However, the resistant strain was only 2.3-fold more resistant to prothiofos than the susceptible strain. The resistant strain included insects having the A298S and G324A mutations in AChE1, which are reportedly involved in prothiofos resistance in P. xylostella, showing reduced AChE sensitivity to inhibition by methamidophos, suggesting that decreased AChE1 sensitivity is one factor conferring acephate resistance. However, allele frequencies at both mutation sites in the resistant strain were low (only 26%). These results suggest that other factors such as GSTs are involved in acephate resistance. Expression of GST genes available in P. xylostella to date was examined using the resistant and susceptible strains, revealing no significant correlation between the expression and resistance levels. 相似文献
14.
为明确河南省部分地区的多花黑麦草Lolium multiflorum种群对乙酰辅酶A羧化酶（acetylCoA carboxylase,ACCase）和乙酰乳酸合成酶（acetolactate synthase,ALS）抑制剂类除草剂的抗性水平和抗性机理,采用整株生物测定法测定采自新乡市和驻马店市的多花黑麦草种群对ACCase抑制剂类除草剂精噁唑禾草灵、炔草酯、唑啉草酯和ALS抑制剂类除草剂甲基二磺隆、氟唑磺隆、啶磺草胺的抗性水平,并对多花黑麦草ACCase和ALS靶标酶编码基因进行克隆及氨基酸序列比对,分析其靶标抗性机理。结果显示,与多花黑麦草敏感种群HNXX01相比,HNZMD04和HNXX05种群对6种除草剂均产生了抗性,HNZMD04种群对精噁唑禾草灵和啶磺草胺的相对抗性倍数分别为44.65和40.31,对炔草酯和氟唑磺隆的相对抗性倍数分别为11.91和11.93;HNXX05种群对精噁唑禾草灵和氟唑磺隆的相对抗性倍数分别为27.70和25.67。HNZMD04和HNXX05抗性种群的ACCase基因均发生了D2078G突变,2个种群的突变率分别为55%和70%;HNZMD04... 相似文献
15.
BACKGROUND: Neonicotinoid action as well as resistance involves interaction with nicotinic acetylcholine receptors (nAChRs). In the housefly, neonicotinoid resistance also involves cytochrome P450, as indicated by bioassay with synergist as well as altered expression. In bioassay, synergism was only partial and indicated possible target‐site resistance. The nAChR α2 subunit is important in neonicotinoid toxicity to insects, and gene expression of the Mdα2 subunit was investigated in field populations and laboratory strains of neonicotinoid‐resistant and insecticide‐susceptible houseflies, Musca domestica L. The genomic sequence covering exon III–VII of Mdα2 was analysed for mutations. RESULTS: Gene expression profiling of Mdα2 revealed notable differences between neonicotinoid‐resistant and insecticide‐susceptible houseflies. On average, the neonicotinoid‐resistant field population 766b and the imidacloprid selected strain 791imi had 60% lower copy numbers of Mdα2 compared with the susceptible reference strain. Sequencing of exon III–VII of the Mdα2, encoding acetylcholine binding‐site regions and three out of four transmembrane domains, did not reveal any mutations explaining the increased neonicotinoid tolerance in the strains examined. CONCLUSION: Previous discoveries and the results of this study suggest that the neonicotinoid resistance mechanism in Danish houseflies involves both cytochrome P450 monooxygenase‐mediated detoxification and reduced expression of the nAChR subunit α2. Copyright © 2010 Society of Chemical Industry 相似文献
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Resistance to chlorpyriphos insecticide in Bemisia tabaci from a field population collected in Benin, West Africa was suggested with bioassay showing the presence of two sub-populations. Patterns of acetylcholinesterase (AChE) inhibition by the organophosphate chlorpyriphos-oxon were analyzed to estimate the number of possible genotypes with different sensitivity expected in three B. tabaci field populations collected in Benin. The analysis of inhibition patterns in these populations compared with four laboratory strains of B. tabaci using chlorpyriphos-oxon allowed the differentiation of three possible genotypes. In the reference strain SUD-S we detected two different acetylcholinesterases with different sensitivity to chlorpyriphos oxon suggesting the presence of two genes ace 1 and ace 2. The proportion of the insensitive enzyme (ace 2) was estimated to be 31%. In field populations we can detect two alleles at the same gene locus ace 1: one susceptible ace1S and one resistant ace1R. Both strains called Arizona University and Mexico-S2 have lost sensitive ace1S but the field populations from Benin clearly contained at least three genotypes confirming heterogeneous populations not completely resistant. 相似文献
18.
Ali Alizadeh Khalil Talebi Vahid Hosseininaveh Mohammad Ghadamyari 《Pesticide biochemistry and physiology》2011,101(2):59-64
The common pistachio psyllid, Agonoscena pistaciae, is the most damaging pest of pistachio in Iran, and is generally controlled by insecticides belonging to various classes especially, phosalone. The toxicity of phosalone in nine populations of the pest was assayed using the residual contact vial and insect-dip methods. The bioassay results showed significant discrepancy in susceptibility to phosalone among the populations. Resistance ratio of the populations to the susceptible population ranged from 3.3 to 11.3. The synergistic effects of TPP, PBO and DEM were evaluated on the susceptible and the most resistant population to determine the involvement of esterases, mixed function oxidases and glutathione S-transferases in resistance mechanisms, respectively. The level of resistance to phosalone in the resistant population was suppressed by TPP, PBO and DEM, suggesting that the resistance to phosalone is mainly caused by esterase detoxification. Biochemical enzyme assays revealed that esterase, glutathione S-transferase and cytochrome P450 monooxygenase activities in the resistant population was higher than that in the susceptible. Glutathione-S-transferases play a minor role in the resistance of the pest to phosalone. 相似文献
19.
BACKGROUND: Bait-formulated spinosad is currently being introduced for housefly (Musca domestica L.) control around the world. Spinosad resistance was evaluated in a multiresistant field population and strains derived from this by selection with insecticides. Constitutive and spinosad-induced expression levels of three cytochrome P450 genes, CYP6A1, CYP6D1 and CYP6D3, previously reported to be involved in insecticide resistance, were examined. RESULTS: In 2004 a baseline for spinosad toxicity of Danish houseflies where all field populations were considered to be susceptible was established. In the present study, females of a multiresistant field population 791a were, however, 27-fold spinosad resistant at LC50, whereas 791a male houseflies were susceptible. Strain 791a was selected with spinosad, thiamethoxam, fipronil and imidacloprid, resulting in four strains with individual characteristics. Selection of 791a with spinosad did not alter spinosad resistance in either males or females, but counterselected against resistance to the insecticides thiamethoxam and imidacloprid targeting nicotinic acetylcholine receptors. A synergist study with piperonyl butoxide, as well as gene expression studies of CYP6A1, CYP6D1 and CYP6D3, indicated a partial involvement of cytochrome P450 genes in spinosad resistance. CONCLUSION: This study reports female-linked spinosad resistance in Danish houseflies. Negative cross-resistance was observed between spinosad and neonicotinoids in one multiresistant housefly strain. Spinosad resistance involved alterations of cytochrome P450 gene expression. Copyright © 2011 Society of Chemical Industry 相似文献
20.
Sameh Boukef Bruce A. McDonald Amor Yahyaoui Salah Rezgui Patrick C. Brunner 《European journal of plant pathology / European Foundation for Plant Pathology》2012,132(1):111-122
The occurrence of fungicide resistance in Mycosphaerella graminicola populations from Tunisia was investigated by examining mutations known to be associated with strobilurin and azole resistance.
Few mutations associated with fungicide resistance were detected. No evidence for strobilurin resistance was found among 357
Tunisian isolates and only two among 80 sequenced isolates carried mutations associated with azole resistance. A network analysis
suggested that these mutations emerged independently from mutations found in previously described European populations. The
population genetic structure of M. graminicola in Tunisia was analyzed using variation at 11 microsatellite loci. Populations in Tunisia were characterized by high gene
and genotype diversity. All populations were in gametic equilibrium and mating type proportions did not deviate from the 1:1
ratio expected under random mating, consistent with regular cycles of sexual reproduction. In combination with a high degree
of gene flow among sampling sites, M. graminicola must be considered a pathogens with high evolutionary potential. Thus, control strategies against Septoria blotch in Tunisia
should be optimized to reduce the emergence and spread of resistant isolates. 相似文献